Monitoring gene expression changes in bovine oviduct epithelial cells during the oestrous cycle
Monitoring gene expression changes in bovine oviduct epithelial cells during the oestrous cycle
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DOI：
10.1677/jme.0.0320449
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发表时间：
2004-04-01
影响因子：
3.5
通讯作者：
Wolf, E
中科院分区：
文献类型：
--
作者：
Bauersachs, S;Rehfeld, S;Wolf, E
输卵管上皮在发情周期期间经历显着的形态和功能变化。为了研究转录组水平上的这些变化，我们结合消减cDNA文库和cDNA阵列杂交，对发情期和间情期的牛输卵管上皮细胞进行了系统的基因表达分析。通过使用源自六只周期性小母牛的 cDNA 探针进行阵列杂交，分析了两个消减文库的总共 3072 个 cDNA 克隆，其中三只在发情期屠宰，三只在间情期屠宰。表达水平显着差异的 cDNA 测序揭示了 77 种不同的 cDNA。 37个基因在发情期表达水平较高，其他40个基因在发情期表达水平较高。已识别的基因代表了多种功能类别。在发情期间，特别是涉及蛋白质分泌和蛋白质修饰调节的基因以及分泌蛋白质的mRNA上调，而在间情期间，特别是涉及转录调节的基因的转录本显示出轻微的上调。通过实时 RT-PCR 对 7 个选定转录本的浓度进行定量，以验证 cDNA 阵列杂交数据。对于所有七个转录本，RT-PCR 结果与阵列杂交获得的结果具有极好的相关性 (r>0.92)。我们的研究首次分析了发情周期不同阶段牛输卵管上皮细胞基因表达谱的变化，为阐明这些细胞的关键转录组变化提供了起点。
The oviduct epithelium undergoes marked morphological and functional changes during the oestrous cycle. To study these changes at the level of the transcriptome we did a systematic gene expression analysis of bovine oviduct epithelial cells at oestrus and dioestrus using a combination of subtracted cDNA libraries and cDNA array hybridisation. A total of 3072 cDNA clones of two subtracted libraries were analysed by array hybridisation with cDNA probes derived from six cyclic heifers, three of them slaughtered at oestrus and three at dioestrus. Sequencing of cDNAs showing significant differences in their expression levels revealed 77 different cDNAs. Thirty-seven were expressed at a higher level at oestrus, for the other 40 genes expression levels were higher at dioestrus. The identified genes represented a variety of functional classes. During oestrus especially genes involved in the regulation of protein secretion and protein modification, and mRNAs of secreted proteins, were up-regulated, whereas during dioestrus particularly transcripts of genes involved in transcription regulation showed a slight up-regulation. The concentrations of seven selected transcripts were quantified by real-time RT-PCR to validate the cDNA array hybridisation data. For all seven transcripts, RT-PCR results were in excellent correlation (r>0.92) with the results obtained by array hybridisation. Our study is the first to analyse changes in gene expression profiles of bovine oviduct epithelial cells during different stages of the oestrous cycle, providing a starting point for the clarification of the key transcriptome changes in these cells.